The human TRAM1 locus expresses circular RNAs

Abstract Numerous indirect and in silico produced evidences suggest circular RNAs (circRNA) in mammals while thorough experimental proofs of their existence have rarely been reported. Biological studies of circRNA, however, should be based on experimentally verified circRNAs. Here, we describe the i...

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Autores principales:	Josephine Dubois, Georg Sczakiel
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/99c3be04329a4556ac01ef5622f903d7
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spelling	oai:doaj.org-article:99c3be04329a4556ac01ef5622f903d72021-11-14T12:23:11ZThe human TRAM1 locus expresses circular RNAs10.1038/s41598-021-01548-02045-2322https://doaj.org/article/99c3be04329a4556ac01ef5622f903d72021-11-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-01548-0https://doaj.org/toc/2045-2322Abstract Numerous indirect and in silico produced evidences suggest circular RNAs (circRNA) in mammals while thorough experimental proofs of their existence have rarely been reported. Biological studies of circRNA, however, should be based on experimentally verified circRNAs. Here, we describe the identification of two circRNAs originating from the gene locus of the translocation associated membrane protein 1 (TRAM1). Linear and potentially circular TRAM1-specific transcripts were identified in a transcriptome analysis of urine RNA of bladder cancer (BCa) patients versus healthy donors. Thus, we first focused on the topology of TRAM1-specific transcripts. We describe conclusive experimental evidence for the existence of TRAM1-specific circRNAs in the human BCa cell lines ECV-304 and RT-4. PCR-based methodology followed by cloning and sequencing strongly indicated the circular topology of two TRAM1 RNAs. Further, studies with exon fusion sequence-specific antisense oligonucleotides (asON) and RNase H as well as studies in the use of RNase R contribute to conclusive set of experiments supporting the circular topology of TRAM1 transcripts. On the biological side, TRAM1-specific circRNAs showed low expression levels and minor differences in BCa cell lines while linear TRAM1 transcripts displayed down-regulated expression in the higher cancer stage model ECV-304 versus more differentiated RT-4 cells.Josephine DuboisGeorg SczakielNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-15 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Josephine Dubois Georg Sczakiel The human TRAM1 locus expresses circular RNAs
description	Abstract Numerous indirect and in silico produced evidences suggest circular RNAs (circRNA) in mammals while thorough experimental proofs of their existence have rarely been reported. Biological studies of circRNA, however, should be based on experimentally verified circRNAs. Here, we describe the identification of two circRNAs originating from the gene locus of the translocation associated membrane protein 1 (TRAM1). Linear and potentially circular TRAM1-specific transcripts were identified in a transcriptome analysis of urine RNA of bladder cancer (BCa) patients versus healthy donors. Thus, we first focused on the topology of TRAM1-specific transcripts. We describe conclusive experimental evidence for the existence of TRAM1-specific circRNAs in the human BCa cell lines ECV-304 and RT-4. PCR-based methodology followed by cloning and sequencing strongly indicated the circular topology of two TRAM1 RNAs. Further, studies with exon fusion sequence-specific antisense oligonucleotides (asON) and RNase H as well as studies in the use of RNase R contribute to conclusive set of experiments supporting the circular topology of TRAM1 transcripts. On the biological side, TRAM1-specific circRNAs showed low expression levels and minor differences in BCa cell lines while linear TRAM1 transcripts displayed down-regulated expression in the higher cancer stage model ECV-304 versus more differentiated RT-4 cells.
format	article
author	Josephine Dubois Georg Sczakiel
author_facet	Josephine Dubois Georg Sczakiel
author_sort	Josephine Dubois
title	The human TRAM1 locus expresses circular RNAs
title_short	The human TRAM1 locus expresses circular RNAs
title_full	The human TRAM1 locus expresses circular RNAs
title_fullStr	The human TRAM1 locus expresses circular RNAs
title_full_unstemmed	The human TRAM1 locus expresses circular RNAs
title_sort	human tram1 locus expresses circular rnas
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/99c3be04329a4556ac01ef5622f903d7
work_keys_str_mv	AT josephinedubois thehumantram1locusexpressescircularrnas AT georgsczakiel thehumantram1locusexpressescircularrnas AT josephinedubois humantram1locusexpressescircularrnas AT georgsczakiel humantram1locusexpressescircularrnas
_version_	1718429245033676800

The human TRAM1 locus expresses circular RNAs

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